Abstract

We demonstrate how the resolution and imaging depth limitations of nonlinear optical microscopy can be overcome by modulating the spatial overlap between two-color pulses. We suppress out-of-focus signals, which limit the imaging depth, by a factor of 100, and enhance the lateral and axial resolution by factors of 1.6 and 1.4–1.8 respectively. Using spatial overlap modulation, we demonstrate background-free three-dimensional imaging of fixed mouse brain tissue at depths for which the signals of the conventional technique are swamped by background noise from out-of-focus regions.

© 2012 OSA

Full Article  |  PDF Article

References

  • View by:
  • |
  • |
  • |

  1. W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science248(4951), 73–76 (1990).
    [CrossRef] [PubMed]
  2. K. König, “Multiphoton microscopy in life sciences,” J. Microsc.200(2), 83–104 (2000).
    [CrossRef] [PubMed]
  3. W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol.21(11), 1369–1377 (2003).
    [CrossRef] [PubMed]
  4. P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nat. Biotechnol.21(11), 1356–1360 (2003).
    [CrossRef] [PubMed]
  5. F. Légaré, C. Pfeffer, and B. R. Olsen, “The role of backscattering in SHG tissue imaging,” Biophys. J.93(4), 1312–1320 (2007).
    [CrossRef] [PubMed]
  6. J. Squier and M. Müller, “High resolution nonlinear microscopy: A review of sources and methods for achieving optimal imaging,” Rev. Sci. Instrum.72(7), 2855–2867 (2001).
    [CrossRef]
  7. Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, “Nonlinear scanning laser microscopy by third harmonic generation,” Appl. Phys. Lett.70(8), 922–924 (1997).
    [CrossRef]
  8. M. D. Duncan, J. Reintjes, and T. J. Manuccia, “Scanning coherent anti-Stokes Raman microscope,” Opt. Lett.7(8), 350–352 (1982).
    [CrossRef] [PubMed]
  9. A. Zumbusch, G. R. Holtom, and X. S. Xie, “Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering,” Phys. Rev. Lett.82(20), 4142–4145 (1999).
    [CrossRef]
  10. K. Isobe, S. Kataoka, R. Murase, W. Watanabe, T. Higashi, S. Kawakami, S. Matsunaga, K. Fukui, and K. Itoh, “Stimulated parametric emission microscopy,” Opt. Express14(2), 786–793 (2006).
    [CrossRef] [PubMed]
  11. K. Isobe, T. Kawasumi, T. Tamaki, S. Kataoka, Y. Ozeki, and K. Itoh, “Three-dimensional profiling of refractive index distribution inside transparent materials by use of nonresonant four-wave mixing microscopy,” Appl. Phys. Express1, 022006 (2008).
    [CrossRef]
  12. W. S. Warren, M. C. Fischer, and T. Ye, “Novel nonlinear contrast improves deep-tissue microscopy,” Laser Focus World43, 99–103 (June 1, 2007).
  13. C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science322(5909), 1857–1861 (2008).
    [CrossRef] [PubMed]
  14. P. Nandakumar, A. Kovalev, and A. Volkmer, “Vibrational imaging based on stimulated Raman scattering microscopy,” New J. Phys.11(3), 033026 (2009).
    [CrossRef]
  15. Y. Ozeki, F. Dake, S. Kajiyama, K. Fukui, and K. Itoh, “Analysis and experimental assessment of the sensitivity of stimulated Raman scattering microscopy,” Opt. Express17(5), 3651–3658 (2009).
    [CrossRef] [PubMed]
  16. P. Tian and W. S. Warren, “Ultrafast measurement of two-photon absorption by loss modulation,” Opt. Lett.27(18), 1634–1636 (2002).
    [CrossRef] [PubMed]
  17. P. Theer, M. T. Hasan, and W. Denk, “Two-photon imaging to a depth of 1000 μm in living brains by use of a Ti:Al2O3 regenerative amplifier,” Opt. Lett.28(12), 1022–1024 (2003).
    [CrossRef] [PubMed]
  18. D. Kobat, M. E. Durst, N. Nishimura, A. W. Wong, C. B. Schaffer, and C. Xu, “Deep tissue multiphoton microscopy using longer wavelength excitation,” Opt. Express17(16), 13354–13364 (2009).
    [CrossRef] [PubMed]
  19. P. Theer and W. Denk, “On the fundamental imaging-depth limit in two-photon microscopy,” J. Opt. Soc. Am. A23(12), 3139–3149 (2006).
    [CrossRef] [PubMed]
  20. G. Zhu, J. van Howe, M. Durst, W. Zipfel, and C. Xu, “Simultaneous spatial and temporal focusing of femtosecond pulses,” Opt. Express13(6), 2153–2159 (2005).
    [CrossRef] [PubMed]
  21. D. Oron, E. Tal, and Y. Silberberg, “Scanningless depth-resolved microscopy,” Opt. Express13(5), 1468–1476 (2005).
    [CrossRef] [PubMed]
  22. A. Leray and J. Mertz, “Rejection of two-photon fluorescence background in thick tissue by differential aberration imaging,” Opt. Express14(22), 10565–10573 (2006).
    [CrossRef] [PubMed]
  23. N. Ji, D. E. Milkie, and E. Betzig, “Adaptive optics via pupil segmentation for high-resolution imaging in biological tissues,” Nat. Methods7(2), 141–147 (2010).
    [CrossRef] [PubMed]
  24. M. A. A. Neil, R. Juskaitis, and T. Wilson, “Method of obtaining optical sectioning by using structured light in a conventional microscope,” Opt. Lett.22(24), 1905–1907 (1997).
    [CrossRef] [PubMed]
  25. N. Chen, C.-H. Wong, and C. J. R. Sheppard, “Focal modulation microscopy,” Opt. Express16(23), 18764–18769 (2008).
    [CrossRef] [PubMed]
  26. S. W. Hell and J. Wichmann, “Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy,” Opt. Lett.19(11), 780–782 (1994).
    [CrossRef] [PubMed]
  27. E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging intracellular fluorescent proteins at nanometer resolution,” Science313(5793), 1642–1645 (2006).
    [CrossRef] [PubMed]
  28. S. T. Hess, T. P. K. Girirajan, and M. D. Mason, “Ultra-high resolution imaging by fluorescence photoactivation localization microscopy,” Biophys. J.91(11), 4258–4272 (2006).
    [CrossRef] [PubMed]
  29. M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods3(10), 793–796 (2006).
    [CrossRef] [PubMed]
  30. K. Fujita, M. Kobayashi, S. Kawano, M. Yamanaka, and S. Kawata, “High-resolution confocal microscopy by saturated excitation of fluorescence,” Phys. Rev. Lett.99(22), 228105 (2007).
    [CrossRef] [PubMed]
  31. K. Isobe, A. Suda, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “High-resolution fluorescence microscopy based on a cyclic sequential multiphoton process,” Biomed. Opt. Express1(3), 791–797 (2010).
    [CrossRef] [PubMed]
  32. M. G. L. Gustafsson, “Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy,” J. Microsc.198(2), 82–87 (2000).
    [CrossRef] [PubMed]
  33. G. Moneron and S. W. Hell, “Two-photon excitation STED microscopy,” Opt. Express17(17), 14567–14573 (2009).
    [CrossRef] [PubMed]
  34. P. E. Hänninen, S. W. Hell, J. Salo, E. Soini, and C. Cremer, “Two-photon excitation 4Pi confocal microscope: Enhanced axial resolution microscope for biological research,” Appl. Phys. Lett.66(13), 1698–1700 (1995).
    [CrossRef]
  35. M. R. Beversluis and S. J. Stranick, “Enhanced contrast coherent anti-Stokes Raman scattering microscopy using annular phase masks,” Appl. Phys. Lett.93(23), 231115 (2008).
    [CrossRef]
  36. R. Boyd, Nonlinear Optics, 3rd ed. (Academic, 2008).
  37. G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
    [CrossRef] [PubMed]
  38. K. Isobe, A. Suda, M. Tanaka, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Nonlinear optical microscopy and spectroscopy employing octave spanning pulses,” IEEE J. Sel. Top. Quantum Electron.16(4), 767–780 (2010).
    [CrossRef]
  39. N. T. Urban, K. I. Willig, S. W. Hell, and U. V. Nägerl, “STED nanoscopy of actin dynamics in synapses deep inside living brain slices,” Biophys. J.101(5), 1277–1284 (2011).
    [CrossRef] [PubMed]
  40. S. Lu, W. Min, S. Chong, G. R. Holtom, and X. S. Xie, “Label-free imaging of heme proteins with two-photon excited photothermal lens microscopy,” Appl. Phys. Lett.96(11), 113701 (2010).
    [CrossRef]

2011 (1)

N. T. Urban, K. I. Willig, S. W. Hell, and U. V. Nägerl, “STED nanoscopy of actin dynamics in synapses deep inside living brain slices,” Biophys. J.101(5), 1277–1284 (2011).
[CrossRef] [PubMed]

2010 (4)

S. Lu, W. Min, S. Chong, G. R. Holtom, and X. S. Xie, “Label-free imaging of heme proteins with two-photon excited photothermal lens microscopy,” Appl. Phys. Lett.96(11), 113701 (2010).
[CrossRef]

K. Isobe, A. Suda, M. Tanaka, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Nonlinear optical microscopy and spectroscopy employing octave spanning pulses,” IEEE J. Sel. Top. Quantum Electron.16(4), 767–780 (2010).
[CrossRef]

N. Ji, D. E. Milkie, and E. Betzig, “Adaptive optics via pupil segmentation for high-resolution imaging in biological tissues,” Nat. Methods7(2), 141–147 (2010).
[CrossRef] [PubMed]

K. Isobe, A. Suda, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “High-resolution fluorescence microscopy based on a cyclic sequential multiphoton process,” Biomed. Opt. Express1(3), 791–797 (2010).
[CrossRef] [PubMed]

2009 (4)

2008 (4)

N. Chen, C.-H. Wong, and C. J. R. Sheppard, “Focal modulation microscopy,” Opt. Express16(23), 18764–18769 (2008).
[CrossRef] [PubMed]

M. R. Beversluis and S. J. Stranick, “Enhanced contrast coherent anti-Stokes Raman scattering microscopy using annular phase masks,” Appl. Phys. Lett.93(23), 231115 (2008).
[CrossRef]

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science322(5909), 1857–1861 (2008).
[CrossRef] [PubMed]

K. Isobe, T. Kawasumi, T. Tamaki, S. Kataoka, Y. Ozeki, and K. Itoh, “Three-dimensional profiling of refractive index distribution inside transparent materials by use of nonresonant four-wave mixing microscopy,” Appl. Phys. Express1, 022006 (2008).
[CrossRef]

2007 (3)

W. S. Warren, M. C. Fischer, and T. Ye, “Novel nonlinear contrast improves deep-tissue microscopy,” Laser Focus World43, 99–103 (June 1, 2007).

F. Légaré, C. Pfeffer, and B. R. Olsen, “The role of backscattering in SHG tissue imaging,” Biophys. J.93(4), 1312–1320 (2007).
[CrossRef] [PubMed]

K. Fujita, M. Kobayashi, S. Kawano, M. Yamanaka, and S. Kawata, “High-resolution confocal microscopy by saturated excitation of fluorescence,” Phys. Rev. Lett.99(22), 228105 (2007).
[CrossRef] [PubMed]

2006 (6)

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging intracellular fluorescent proteins at nanometer resolution,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

S. T. Hess, T. P. K. Girirajan, and M. D. Mason, “Ultra-high resolution imaging by fluorescence photoactivation localization microscopy,” Biophys. J.91(11), 4258–4272 (2006).
[CrossRef] [PubMed]

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods3(10), 793–796 (2006).
[CrossRef] [PubMed]

K. Isobe, S. Kataoka, R. Murase, W. Watanabe, T. Higashi, S. Kawakami, S. Matsunaga, K. Fukui, and K. Itoh, “Stimulated parametric emission microscopy,” Opt. Express14(2), 786–793 (2006).
[CrossRef] [PubMed]

A. Leray and J. Mertz, “Rejection of two-photon fluorescence background in thick tissue by differential aberration imaging,” Opt. Express14(22), 10565–10573 (2006).
[CrossRef] [PubMed]

P. Theer and W. Denk, “On the fundamental imaging-depth limit in two-photon microscopy,” J. Opt. Soc. Am. A23(12), 3139–3149 (2006).
[CrossRef] [PubMed]

2005 (2)

2003 (3)

P. Theer, M. T. Hasan, and W. Denk, “Two-photon imaging to a depth of 1000 μm in living brains by use of a Ti:Al2O3 regenerative amplifier,” Opt. Lett.28(12), 1022–1024 (2003).
[CrossRef] [PubMed]

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol.21(11), 1369–1377 (2003).
[CrossRef] [PubMed]

P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nat. Biotechnol.21(11), 1356–1360 (2003).
[CrossRef] [PubMed]

2002 (1)

2001 (1)

J. Squier and M. Müller, “High resolution nonlinear microscopy: A review of sources and methods for achieving optimal imaging,” Rev. Sci. Instrum.72(7), 2855–2867 (2001).
[CrossRef]

2000 (3)

K. König, “Multiphoton microscopy in life sciences,” J. Microsc.200(2), 83–104 (2000).
[CrossRef] [PubMed]

M. G. L. Gustafsson, “Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy,” J. Microsc.198(2), 82–87 (2000).
[CrossRef] [PubMed]

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
[CrossRef] [PubMed]

1999 (1)

A. Zumbusch, G. R. Holtom, and X. S. Xie, “Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering,” Phys. Rev. Lett.82(20), 4142–4145 (1999).
[CrossRef]

1997 (2)

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, “Nonlinear scanning laser microscopy by third harmonic generation,” Appl. Phys. Lett.70(8), 922–924 (1997).
[CrossRef]

M. A. A. Neil, R. Juskaitis, and T. Wilson, “Method of obtaining optical sectioning by using structured light in a conventional microscope,” Opt. Lett.22(24), 1905–1907 (1997).
[CrossRef] [PubMed]

1995 (1)

P. E. Hänninen, S. W. Hell, J. Salo, E. Soini, and C. Cremer, “Two-photon excitation 4Pi confocal microscope: Enhanced axial resolution microscope for biological research,” Appl. Phys. Lett.66(13), 1698–1700 (1995).
[CrossRef]

1994 (1)

1990 (1)

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science248(4951), 73–76 (1990).
[CrossRef] [PubMed]

1982 (1)

Barad, Y.

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, “Nonlinear scanning laser microscopy by third harmonic generation,” Appl. Phys. Lett.70(8), 922–924 (1997).
[CrossRef]

Bates, M.

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods3(10), 793–796 (2006).
[CrossRef] [PubMed]

Bernstein, M.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
[CrossRef] [PubMed]

Betzig, E.

N. Ji, D. E. Milkie, and E. Betzig, “Adaptive optics via pupil segmentation for high-resolution imaging in biological tissues,” Nat. Methods7(2), 141–147 (2010).
[CrossRef] [PubMed]

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging intracellular fluorescent proteins at nanometer resolution,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Beversluis, M. R.

M. R. Beversluis and S. J. Stranick, “Enhanced contrast coherent anti-Stokes Raman scattering microscopy using annular phase masks,” Appl. Phys. Lett.93(23), 231115 (2008).
[CrossRef]

Bonifacino, J. S.

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging intracellular fluorescent proteins at nanometer resolution,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Campagnola, P. J.

P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nat. Biotechnol.21(11), 1356–1360 (2003).
[CrossRef] [PubMed]

Chen, N.

Chong, S.

S. Lu, W. Min, S. Chong, G. R. Holtom, and X. S. Xie, “Label-free imaging of heme proteins with two-photon excited photothermal lens microscopy,” Appl. Phys. Lett.96(11), 113701 (2010).
[CrossRef]

Cremer, C.

P. E. Hänninen, S. W. Hell, J. Salo, E. Soini, and C. Cremer, “Two-photon excitation 4Pi confocal microscope: Enhanced axial resolution microscope for biological research,” Appl. Phys. Lett.66(13), 1698–1700 (1995).
[CrossRef]

Dake, F.

Davidson, M. W.

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging intracellular fluorescent proteins at nanometer resolution,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Denk, W.

Duncan, M. D.

Durst, M.

Durst, M. E.

Eisenberg, H.

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, “Nonlinear scanning laser microscopy by third harmonic generation,” Appl. Phys. Lett.70(8), 922–924 (1997).
[CrossRef]

Feng, G.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
[CrossRef] [PubMed]

Fischer, M. C.

W. S. Warren, M. C. Fischer, and T. Ye, “Novel nonlinear contrast improves deep-tissue microscopy,” Laser Focus World43, 99–103 (June 1, 2007).

Freudiger, C. W.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science322(5909), 1857–1861 (2008).
[CrossRef] [PubMed]

Fujita, K.

K. Fujita, M. Kobayashi, S. Kawano, M. Yamanaka, and S. Kawata, “High-resolution confocal microscopy by saturated excitation of fluorescence,” Phys. Rev. Lett.99(22), 228105 (2007).
[CrossRef] [PubMed]

Fukui, K.

Girirajan, T. P. K.

S. T. Hess, T. P. K. Girirajan, and M. D. Mason, “Ultra-high resolution imaging by fluorescence photoactivation localization microscopy,” Biophys. J.91(11), 4258–4272 (2006).
[CrossRef] [PubMed]

Gustafsson, M. G. L.

M. G. L. Gustafsson, “Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy,” J. Microsc.198(2), 82–87 (2000).
[CrossRef] [PubMed]

Hänninen, P. E.

P. E. Hänninen, S. W. Hell, J. Salo, E. Soini, and C. Cremer, “Two-photon excitation 4Pi confocal microscope: Enhanced axial resolution microscope for biological research,” Appl. Phys. Lett.66(13), 1698–1700 (1995).
[CrossRef]

Hasan, M. T.

Hashimoto, H.

K. Isobe, A. Suda, M. Tanaka, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Nonlinear optical microscopy and spectroscopy employing octave spanning pulses,” IEEE J. Sel. Top. Quantum Electron.16(4), 767–780 (2010).
[CrossRef]

K. Isobe, A. Suda, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “High-resolution fluorescence microscopy based on a cyclic sequential multiphoton process,” Biomed. Opt. Express1(3), 791–797 (2010).
[CrossRef] [PubMed]

He, C.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science322(5909), 1857–1861 (2008).
[CrossRef] [PubMed]

Hell, S. W.

N. T. Urban, K. I. Willig, S. W. Hell, and U. V. Nägerl, “STED nanoscopy of actin dynamics in synapses deep inside living brain slices,” Biophys. J.101(5), 1277–1284 (2011).
[CrossRef] [PubMed]

G. Moneron and S. W. Hell, “Two-photon excitation STED microscopy,” Opt. Express17(17), 14567–14573 (2009).
[CrossRef] [PubMed]

P. E. Hänninen, S. W. Hell, J. Salo, E. Soini, and C. Cremer, “Two-photon excitation 4Pi confocal microscope: Enhanced axial resolution microscope for biological research,” Appl. Phys. Lett.66(13), 1698–1700 (1995).
[CrossRef]

S. W. Hell and J. Wichmann, “Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy,” Opt. Lett.19(11), 780–782 (1994).
[CrossRef] [PubMed]

Hess, H. F.

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging intracellular fluorescent proteins at nanometer resolution,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Hess, S. T.

S. T. Hess, T. P. K. Girirajan, and M. D. Mason, “Ultra-high resolution imaging by fluorescence photoactivation localization microscopy,” Biophys. J.91(11), 4258–4272 (2006).
[CrossRef] [PubMed]

Higashi, T.

Holtom, G. R.

S. Lu, W. Min, S. Chong, G. R. Holtom, and X. S. Xie, “Label-free imaging of heme proteins with two-photon excited photothermal lens microscopy,” Appl. Phys. Lett.96(11), 113701 (2010).
[CrossRef]

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science322(5909), 1857–1861 (2008).
[CrossRef] [PubMed]

A. Zumbusch, G. R. Holtom, and X. S. Xie, “Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering,” Phys. Rev. Lett.82(20), 4142–4145 (1999).
[CrossRef]

Horowitz, M.

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, “Nonlinear scanning laser microscopy by third harmonic generation,” Appl. Phys. Lett.70(8), 922–924 (1997).
[CrossRef]

Isobe, K.

K. Isobe, A. Suda, M. Tanaka, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Nonlinear optical microscopy and spectroscopy employing octave spanning pulses,” IEEE J. Sel. Top. Quantum Electron.16(4), 767–780 (2010).
[CrossRef]

K. Isobe, A. Suda, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “High-resolution fluorescence microscopy based on a cyclic sequential multiphoton process,” Biomed. Opt. Express1(3), 791–797 (2010).
[CrossRef] [PubMed]

K. Isobe, T. Kawasumi, T. Tamaki, S. Kataoka, Y. Ozeki, and K. Itoh, “Three-dimensional profiling of refractive index distribution inside transparent materials by use of nonresonant four-wave mixing microscopy,” Appl. Phys. Express1, 022006 (2008).
[CrossRef]

K. Isobe, S. Kataoka, R. Murase, W. Watanabe, T. Higashi, S. Kawakami, S. Matsunaga, K. Fukui, and K. Itoh, “Stimulated parametric emission microscopy,” Opt. Express14(2), 786–793 (2006).
[CrossRef] [PubMed]

Itoh, K.

Ji, N.

N. Ji, D. E. Milkie, and E. Betzig, “Adaptive optics via pupil segmentation for high-resolution imaging in biological tissues,” Nat. Methods7(2), 141–147 (2010).
[CrossRef] [PubMed]

Juskaitis, R.

Kajiyama, S.

Kang, J. X.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science322(5909), 1857–1861 (2008).
[CrossRef] [PubMed]

Kannari, F.

K. Isobe, A. Suda, M. Tanaka, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Nonlinear optical microscopy and spectroscopy employing octave spanning pulses,” IEEE J. Sel. Top. Quantum Electron.16(4), 767–780 (2010).
[CrossRef]

K. Isobe, A. Suda, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “High-resolution fluorescence microscopy based on a cyclic sequential multiphoton process,” Biomed. Opt. Express1(3), 791–797 (2010).
[CrossRef] [PubMed]

Kataoka, S.

K. Isobe, T. Kawasumi, T. Tamaki, S. Kataoka, Y. Ozeki, and K. Itoh, “Three-dimensional profiling of refractive index distribution inside transparent materials by use of nonresonant four-wave mixing microscopy,” Appl. Phys. Express1, 022006 (2008).
[CrossRef]

K. Isobe, S. Kataoka, R. Murase, W. Watanabe, T. Higashi, S. Kawakami, S. Matsunaga, K. Fukui, and K. Itoh, “Stimulated parametric emission microscopy,” Opt. Express14(2), 786–793 (2006).
[CrossRef] [PubMed]

Kawakami, S.

Kawano, H.

K. Isobe, A. Suda, M. Tanaka, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Nonlinear optical microscopy and spectroscopy employing octave spanning pulses,” IEEE J. Sel. Top. Quantum Electron.16(4), 767–780 (2010).
[CrossRef]

K. Isobe, A. Suda, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “High-resolution fluorescence microscopy based on a cyclic sequential multiphoton process,” Biomed. Opt. Express1(3), 791–797 (2010).
[CrossRef] [PubMed]

Kawano, S.

K. Fujita, M. Kobayashi, S. Kawano, M. Yamanaka, and S. Kawata, “High-resolution confocal microscopy by saturated excitation of fluorescence,” Phys. Rev. Lett.99(22), 228105 (2007).
[CrossRef] [PubMed]

Kawasumi, T.

K. Isobe, T. Kawasumi, T. Tamaki, S. Kataoka, Y. Ozeki, and K. Itoh, “Three-dimensional profiling of refractive index distribution inside transparent materials by use of nonresonant four-wave mixing microscopy,” Appl. Phys. Express1, 022006 (2008).
[CrossRef]

Kawata, S.

K. Fujita, M. Kobayashi, S. Kawano, M. Yamanaka, and S. Kawata, “High-resolution confocal microscopy by saturated excitation of fluorescence,” Phys. Rev. Lett.99(22), 228105 (2007).
[CrossRef] [PubMed]

Keller-Peck, C.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
[CrossRef] [PubMed]

Kobat, D.

Kobayashi, M.

K. Fujita, M. Kobayashi, S. Kawano, M. Yamanaka, and S. Kawata, “High-resolution confocal microscopy by saturated excitation of fluorescence,” Phys. Rev. Lett.99(22), 228105 (2007).
[CrossRef] [PubMed]

König, K.

K. König, “Multiphoton microscopy in life sciences,” J. Microsc.200(2), 83–104 (2000).
[CrossRef] [PubMed]

Kovalev, A.

P. Nandakumar, A. Kovalev, and A. Volkmer, “Vibrational imaging based on stimulated Raman scattering microscopy,” New J. Phys.11(3), 033026 (2009).
[CrossRef]

Légaré, F.

F. Légaré, C. Pfeffer, and B. R. Olsen, “The role of backscattering in SHG tissue imaging,” Biophys. J.93(4), 1312–1320 (2007).
[CrossRef] [PubMed]

Leray, A.

Lichtman, J. W.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
[CrossRef] [PubMed]

Lindwasser, O. W.

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging intracellular fluorescent proteins at nanometer resolution,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Lippincott-Schwartz, J.

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging intracellular fluorescent proteins at nanometer resolution,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Loew, L. M.

P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nat. Biotechnol.21(11), 1356–1360 (2003).
[CrossRef] [PubMed]

Lu, S.

S. Lu, W. Min, S. Chong, G. R. Holtom, and X. S. Xie, “Label-free imaging of heme proteins with two-photon excited photothermal lens microscopy,” Appl. Phys. Lett.96(11), 113701 (2010).
[CrossRef]

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science322(5909), 1857–1861 (2008).
[CrossRef] [PubMed]

Manuccia, T. J.

Mason, M. D.

S. T. Hess, T. P. K. Girirajan, and M. D. Mason, “Ultra-high resolution imaging by fluorescence photoactivation localization microscopy,” Biophys. J.91(11), 4258–4272 (2006).
[CrossRef] [PubMed]

Matsunaga, S.

Mellor, R. H.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
[CrossRef] [PubMed]

Mertz, J.

Midorikawa, K.

K. Isobe, A. Suda, M. Tanaka, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Nonlinear optical microscopy and spectroscopy employing octave spanning pulses,” IEEE J. Sel. Top. Quantum Electron.16(4), 767–780 (2010).
[CrossRef]

K. Isobe, A. Suda, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “High-resolution fluorescence microscopy based on a cyclic sequential multiphoton process,” Biomed. Opt. Express1(3), 791–797 (2010).
[CrossRef] [PubMed]

Milkie, D. E.

N. Ji, D. E. Milkie, and E. Betzig, “Adaptive optics via pupil segmentation for high-resolution imaging in biological tissues,” Nat. Methods7(2), 141–147 (2010).
[CrossRef] [PubMed]

Min, W.

S. Lu, W. Min, S. Chong, G. R. Holtom, and X. S. Xie, “Label-free imaging of heme proteins with two-photon excited photothermal lens microscopy,” Appl. Phys. Lett.96(11), 113701 (2010).
[CrossRef]

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science322(5909), 1857–1861 (2008).
[CrossRef] [PubMed]

Miyawaki, A.

K. Isobe, A. Suda, M. Tanaka, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Nonlinear optical microscopy and spectroscopy employing octave spanning pulses,” IEEE J. Sel. Top. Quantum Electron.16(4), 767–780 (2010).
[CrossRef]

K. Isobe, A. Suda, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “High-resolution fluorescence microscopy based on a cyclic sequential multiphoton process,” Biomed. Opt. Express1(3), 791–797 (2010).
[CrossRef] [PubMed]

Mizuno, H.

K. Isobe, A. Suda, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “High-resolution fluorescence microscopy based on a cyclic sequential multiphoton process,” Biomed. Opt. Express1(3), 791–797 (2010).
[CrossRef] [PubMed]

K. Isobe, A. Suda, M. Tanaka, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Nonlinear optical microscopy and spectroscopy employing octave spanning pulses,” IEEE J. Sel. Top. Quantum Electron.16(4), 767–780 (2010).
[CrossRef]

Moneron, G.

Müller, M.

J. Squier and M. Müller, “High resolution nonlinear microscopy: A review of sources and methods for achieving optimal imaging,” Rev. Sci. Instrum.72(7), 2855–2867 (2001).
[CrossRef]

Murase, R.

Nägerl, U. V.

N. T. Urban, K. I. Willig, S. W. Hell, and U. V. Nägerl, “STED nanoscopy of actin dynamics in synapses deep inside living brain slices,” Biophys. J.101(5), 1277–1284 (2011).
[CrossRef] [PubMed]

Nandakumar, P.

P. Nandakumar, A. Kovalev, and A. Volkmer, “Vibrational imaging based on stimulated Raman scattering microscopy,” New J. Phys.11(3), 033026 (2009).
[CrossRef]

Neil, M. A. A.

Nerbonne, J. M.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
[CrossRef] [PubMed]

Nguyen, Q. T.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
[CrossRef] [PubMed]

Nishimura, N.

Olenych, S.

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging intracellular fluorescent proteins at nanometer resolution,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Olsen, B. R.

F. Légaré, C. Pfeffer, and B. R. Olsen, “The role of backscattering in SHG tissue imaging,” Biophys. J.93(4), 1312–1320 (2007).
[CrossRef] [PubMed]

Oron, D.

Ozeki, Y.

Y. Ozeki, F. Dake, S. Kajiyama, K. Fukui, and K. Itoh, “Analysis and experimental assessment of the sensitivity of stimulated Raman scattering microscopy,” Opt. Express17(5), 3651–3658 (2009).
[CrossRef] [PubMed]

K. Isobe, T. Kawasumi, T. Tamaki, S. Kataoka, Y. Ozeki, and K. Itoh, “Three-dimensional profiling of refractive index distribution inside transparent materials by use of nonresonant four-wave mixing microscopy,” Appl. Phys. Express1, 022006 (2008).
[CrossRef]

Patterson, G. H.

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging intracellular fluorescent proteins at nanometer resolution,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Pfeffer, C.

F. Légaré, C. Pfeffer, and B. R. Olsen, “The role of backscattering in SHG tissue imaging,” Biophys. J.93(4), 1312–1320 (2007).
[CrossRef] [PubMed]

Reintjes, J.

Rust, M. J.

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods3(10), 793–796 (2006).
[CrossRef] [PubMed]

Saar, B. G.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science322(5909), 1857–1861 (2008).
[CrossRef] [PubMed]

Salo, J.

P. E. Hänninen, S. W. Hell, J. Salo, E. Soini, and C. Cremer, “Two-photon excitation 4Pi confocal microscope: Enhanced axial resolution microscope for biological research,” Appl. Phys. Lett.66(13), 1698–1700 (1995).
[CrossRef]

Sanes, J. R.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
[CrossRef] [PubMed]

Schaffer, C. B.

Sheppard, C. J. R.

Silberberg, Y.

D. Oron, E. Tal, and Y. Silberberg, “Scanningless depth-resolved microscopy,” Opt. Express13(5), 1468–1476 (2005).
[CrossRef] [PubMed]

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, “Nonlinear scanning laser microscopy by third harmonic generation,” Appl. Phys. Lett.70(8), 922–924 (1997).
[CrossRef]

Soini, E.

P. E. Hänninen, S. W. Hell, J. Salo, E. Soini, and C. Cremer, “Two-photon excitation 4Pi confocal microscope: Enhanced axial resolution microscope for biological research,” Appl. Phys. Lett.66(13), 1698–1700 (1995).
[CrossRef]

Sougrat, R.

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging intracellular fluorescent proteins at nanometer resolution,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Squier, J.

J. Squier and M. Müller, “High resolution nonlinear microscopy: A review of sources and methods for achieving optimal imaging,” Rev. Sci. Instrum.72(7), 2855–2867 (2001).
[CrossRef]

Stranick, S. J.

M. R. Beversluis and S. J. Stranick, “Enhanced contrast coherent anti-Stokes Raman scattering microscopy using annular phase masks,” Appl. Phys. Lett.93(23), 231115 (2008).
[CrossRef]

Strickler, J. H.

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science248(4951), 73–76 (1990).
[CrossRef] [PubMed]

Suda, A.

K. Isobe, A. Suda, M. Tanaka, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Nonlinear optical microscopy and spectroscopy employing octave spanning pulses,” IEEE J. Sel. Top. Quantum Electron.16(4), 767–780 (2010).
[CrossRef]

K. Isobe, A. Suda, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “High-resolution fluorescence microscopy based on a cyclic sequential multiphoton process,” Biomed. Opt. Express1(3), 791–797 (2010).
[CrossRef] [PubMed]

Tal, E.

Tamaki, T.

K. Isobe, T. Kawasumi, T. Tamaki, S. Kataoka, Y. Ozeki, and K. Itoh, “Three-dimensional profiling of refractive index distribution inside transparent materials by use of nonresonant four-wave mixing microscopy,” Appl. Phys. Express1, 022006 (2008).
[CrossRef]

Tanaka, M.

K. Isobe, A. Suda, M. Tanaka, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Nonlinear optical microscopy and spectroscopy employing octave spanning pulses,” IEEE J. Sel. Top. Quantum Electron.16(4), 767–780 (2010).
[CrossRef]

Theer, P.

Tian, P.

Tsai, J. C.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science322(5909), 1857–1861 (2008).
[CrossRef] [PubMed]

Urban, N. T.

N. T. Urban, K. I. Willig, S. W. Hell, and U. V. Nägerl, “STED nanoscopy of actin dynamics in synapses deep inside living brain slices,” Biophys. J.101(5), 1277–1284 (2011).
[CrossRef] [PubMed]

van Howe, J.

Volkmer, A.

P. Nandakumar, A. Kovalev, and A. Volkmer, “Vibrational imaging based on stimulated Raman scattering microscopy,” New J. Phys.11(3), 033026 (2009).
[CrossRef]

Wallace, M.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
[CrossRef] [PubMed]

Warren, W. S.

W. S. Warren, M. C. Fischer, and T. Ye, “Novel nonlinear contrast improves deep-tissue microscopy,” Laser Focus World43, 99–103 (June 1, 2007).

P. Tian and W. S. Warren, “Ultrafast measurement of two-photon absorption by loss modulation,” Opt. Lett.27(18), 1634–1636 (2002).
[CrossRef] [PubMed]

Watanabe, W.

Webb, W. W.

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol.21(11), 1369–1377 (2003).
[CrossRef] [PubMed]

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science248(4951), 73–76 (1990).
[CrossRef] [PubMed]

Wichmann, J.

Williams, R. M.

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol.21(11), 1369–1377 (2003).
[CrossRef] [PubMed]

Willig, K. I.

N. T. Urban, K. I. Willig, S. W. Hell, and U. V. Nägerl, “STED nanoscopy of actin dynamics in synapses deep inside living brain slices,” Biophys. J.101(5), 1277–1284 (2011).
[CrossRef] [PubMed]

Wilson, T.

Wong, A. W.

Wong, C.-H.

Xie, X. S.

S. Lu, W. Min, S. Chong, G. R. Holtom, and X. S. Xie, “Label-free imaging of heme proteins with two-photon excited photothermal lens microscopy,” Appl. Phys. Lett.96(11), 113701 (2010).
[CrossRef]

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science322(5909), 1857–1861 (2008).
[CrossRef] [PubMed]

A. Zumbusch, G. R. Holtom, and X. S. Xie, “Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering,” Phys. Rev. Lett.82(20), 4142–4145 (1999).
[CrossRef]

Xu, C.

Yamanaka, M.

K. Fujita, M. Kobayashi, S. Kawano, M. Yamanaka, and S. Kawata, “High-resolution confocal microscopy by saturated excitation of fluorescence,” Phys. Rev. Lett.99(22), 228105 (2007).
[CrossRef] [PubMed]

Ye, T.

W. S. Warren, M. C. Fischer, and T. Ye, “Novel nonlinear contrast improves deep-tissue microscopy,” Laser Focus World43, 99–103 (June 1, 2007).

Zhu, G.

Zhuang, X.

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods3(10), 793–796 (2006).
[CrossRef] [PubMed]

Zipfel, W.

Zipfel, W. R.

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol.21(11), 1369–1377 (2003).
[CrossRef] [PubMed]

Zumbusch, A.

A. Zumbusch, G. R. Holtom, and X. S. Xie, “Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering,” Phys. Rev. Lett.82(20), 4142–4145 (1999).
[CrossRef]

Appl. Phys. Express (1)

K. Isobe, T. Kawasumi, T. Tamaki, S. Kataoka, Y. Ozeki, and K. Itoh, “Three-dimensional profiling of refractive index distribution inside transparent materials by use of nonresonant four-wave mixing microscopy,” Appl. Phys. Express1, 022006 (2008).
[CrossRef]

Appl. Phys. Lett. (4)

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, “Nonlinear scanning laser microscopy by third harmonic generation,” Appl. Phys. Lett.70(8), 922–924 (1997).
[CrossRef]

P. E. Hänninen, S. W. Hell, J. Salo, E. Soini, and C. Cremer, “Two-photon excitation 4Pi confocal microscope: Enhanced axial resolution microscope for biological research,” Appl. Phys. Lett.66(13), 1698–1700 (1995).
[CrossRef]

M. R. Beversluis and S. J. Stranick, “Enhanced contrast coherent anti-Stokes Raman scattering microscopy using annular phase masks,” Appl. Phys. Lett.93(23), 231115 (2008).
[CrossRef]

S. Lu, W. Min, S. Chong, G. R. Holtom, and X. S. Xie, “Label-free imaging of heme proteins with two-photon excited photothermal lens microscopy,” Appl. Phys. Lett.96(11), 113701 (2010).
[CrossRef]

Biomed. Opt. Express (1)

Biophys. J. (3)

N. T. Urban, K. I. Willig, S. W. Hell, and U. V. Nägerl, “STED nanoscopy of actin dynamics in synapses deep inside living brain slices,” Biophys. J.101(5), 1277–1284 (2011).
[CrossRef] [PubMed]

S. T. Hess, T. P. K. Girirajan, and M. D. Mason, “Ultra-high resolution imaging by fluorescence photoactivation localization microscopy,” Biophys. J.91(11), 4258–4272 (2006).
[CrossRef] [PubMed]

F. Légaré, C. Pfeffer, and B. R. Olsen, “The role of backscattering in SHG tissue imaging,” Biophys. J.93(4), 1312–1320 (2007).
[CrossRef] [PubMed]

IEEE J. Sel. Top. Quantum Electron. (1)

K. Isobe, A. Suda, M. Tanaka, H. Hashimoto, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Nonlinear optical microscopy and spectroscopy employing octave spanning pulses,” IEEE J. Sel. Top. Quantum Electron.16(4), 767–780 (2010).
[CrossRef]

J. Microsc. (2)

K. König, “Multiphoton microscopy in life sciences,” J. Microsc.200(2), 83–104 (2000).
[CrossRef] [PubMed]

M. G. L. Gustafsson, “Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy,” J. Microsc.198(2), 82–87 (2000).
[CrossRef] [PubMed]

J. Opt. Soc. Am. A (1)

Laser Focus World (1)

W. S. Warren, M. C. Fischer, and T. Ye, “Novel nonlinear contrast improves deep-tissue microscopy,” Laser Focus World43, 99–103 (June 1, 2007).

Nat. Biotechnol. (2)

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol.21(11), 1369–1377 (2003).
[CrossRef] [PubMed]

P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nat. Biotechnol.21(11), 1356–1360 (2003).
[CrossRef] [PubMed]

Nat. Methods (2)

N. Ji, D. E. Milkie, and E. Betzig, “Adaptive optics via pupil segmentation for high-resolution imaging in biological tissues,” Nat. Methods7(2), 141–147 (2010).
[CrossRef] [PubMed]

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods3(10), 793–796 (2006).
[CrossRef] [PubMed]

Neuron (1)

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
[CrossRef] [PubMed]

New J. Phys. (1)

P. Nandakumar, A. Kovalev, and A. Volkmer, “Vibrational imaging based on stimulated Raman scattering microscopy,” New J. Phys.11(3), 033026 (2009).
[CrossRef]

Opt. Express (8)

Opt. Lett. (5)

Phys. Rev. Lett. (2)

A. Zumbusch, G. R. Holtom, and X. S. Xie, “Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering,” Phys. Rev. Lett.82(20), 4142–4145 (1999).
[CrossRef]

K. Fujita, M. Kobayashi, S. Kawano, M. Yamanaka, and S. Kawata, “High-resolution confocal microscopy by saturated excitation of fluorescence,” Phys. Rev. Lett.99(22), 228105 (2007).
[CrossRef] [PubMed]

Rev. Sci. Instrum. (1)

J. Squier and M. Müller, “High resolution nonlinear microscopy: A review of sources and methods for achieving optimal imaging,” Rev. Sci. Instrum.72(7), 2855–2867 (2001).
[CrossRef]

Science (3)

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science322(5909), 1857–1861 (2008).
[CrossRef] [PubMed]

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science248(4951), 73–76 (1990).
[CrossRef] [PubMed]

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging intracellular fluorescent proteins at nanometer resolution,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Other (1)

R. Boyd, Nonlinear Optics, 3rd ed. (Academic, 2008).

Cited By

OSA participates in CrossRef's Cited-By Linking service. Citing articles from OSA journals and other participating publishers are listed here.

Alert me when this article is cited.


Figures (8)

Fig. 1
Fig. 1

Modulation scheme and principle of SPOM-NOM. (a) Beam-pointing modulation. The spatial overlap between the two-color pulses is modulated by moving the focal position of one of the two pulses by beam pointing modulation. (b) Transition of the intensity distributions of the two pulses in SPOM. The focal position of one pulse is moved with simple harmonic motion between –δ and +δ at a frequency of f. (c) Variation of the nondegenerate TPEF intensity by SPOM. The TPEF intensity at position 0 is modulated at a frequency of 2f, while those at positions –δ and +δ are modulated at a frequency of f. By exploiting the different frequency dependences, information in a volume smaller than the interaction volume can be obtained from the demodulation signal at 2f. (d) Input intensities for TPA or SRS detection. (e) Intensity changes by TPA. (f) Intensity changes by SRS.

Fig. 2
Fig. 2

SPOM-NOM setup. OPO: optical parametric oscillator, PCs: pre-chirpers, DMs: dichroic mirrors, OB: objective lens, SPF: short-pass filter, BPF: band-pass filter, PMT: photomultiplier tube.

Fig. 3
Fig. 3

Enhancement of the spatial resolution in TPEF imaging by SPOM-NOM. (a, b) TPEF images of a 100-nm florescent bead by (a) conventional microscopy and (b) SPOM-NOM. (c) TPEF intensity and demodulated intensity profiles in the x direction in images (a) and (b), which corresponds to the lateral response. (d) Superposition of the measured TPEF intensity profile of a 100-nm fluorescent bead and the TPEF intensity profile obtained by shifting it. (e) TPEF intensity and demodulated intensity profiles in the z direction in images (a) and (b), which corresponds to the axial response. (f, g) TPEF images of 200-nm florescent beads obtained by (f) conventional microscopy and (g) SPOM-NOM. The sharper response of SPOM-NOM results in a better spatial separation of the beads. (h) Intensity and demodulated intensity profiles along the dashed lines in (f) and (g).

Fig. 4
Fig. 4

Enhancement of the spatial resolution in SFG imaging by SPOM-NOM. (a, b) SFG images of granulated sugar pounded in a mortar obtained by (a) conventional microscopy and (b) SPOM-NOM. (c) Intensity and demodulated intensity profiles along the dashed lines in (a) and (b). (d) Line profiles of (demodulated) SFG and SHG intensities in the axial direction near the surface of a quartz crystal cut on 90°. (e) The maximum displacement dependence of the demodulated intensity.

Fig. 5
Fig. 5

Suppression of out-of-focus nonlinear signals by the SPOM technique. (a) Line profiles of TPEF intensity and demodulated TPEF intensity in the axial direction near the interface between a cyan-fluorescent protein solution and a glass slide. (b) Intensity distributions of SFG with 775 and 1000-nm pulses, SHG with 775-nm pulses and SHG with 1000-nm pulses, and demodulated SFG distribution along the axial direction near the surface of a quartz crystal cut on 90°.

Fig. 6
Fig. 6

(a) TPEF images of a tissue-like phantom, composed of low-melting-point agarose gel containing 2-μm fluorescent polystyrene beads at a concentration of 1.0 × 109 beads/mL, by conventional microscopy (upper panel) and SPOM-NOM (bottom panel). The background that includes the out-of-focus TPEF signals has been suppressed by the SPOM technique. (b) The contrast ratio between the TPEF intensity of fluorescent beads in the focal plane and the background TPEF intensity as a function of penetration depth in a tissue-like phantom.

Fig. 7
Fig. 7

TPEF images of fixed mouse brain tissues, which express a YFP in a subset of neurons. (a, b) 3D TPEF images obtained by (a) conventional microscopy and (b) SPOM-NOM. Each 3D image was reconstructed from 295 xy images (32 × 64 pixels) obtained at depth increments of 1 μm. (c, d) Maximum-intensity x projections of the image stacks for (c) conventional microscopy and (d) SPOM-NOM in (a) and (b).

Fig. 8
Fig. 8

Sensing loss by SRS with the SPOM technique. (a, b) SRS images at frequency differences of (a) 3051 cm–1 (aromatic C–H stretching vibration) and (b) 3333 cm–1 (nonresonant) by SPOM-NOM. (c, d) CARS images at frequency differences of (c) 3051 cm–1 and (d) 3333 cm–1 by conventional microscopy. The nonresonant signals in the SRS image are suppressed, whereas those in the CARS image are relatively large.

Equations (9)

Equations on this page are rendered with MathJax. Learn more.

S conv (r,z)=η σ (2) 0 T I 1 (r,z,t) I 2 (r,z,t) dt η σ (2) I 1 (peak) (r,z) I 2 (peak) (r,z) f L τT η σ (2) f L τ I 1 (ave) (r,z) I 2 (ave) (r,z)T,
S 2mf (r,z) η σ (2) I 2 (ave) (r,z) f L τ × 0 T I 1 (ave) (rδ(z)cos2πft,z) cos( 2π2mft+mπ )dt,
δ(z)= δ 0 (z+ f OB ) f OB ,
I 1 (ave) (rδ(z)cos2πft)= l=0 1 l! l I 1 (ave) (r,z) r l | r=0 ( rδ(z)cos2πft ) l = l=0 1 l! l I 1 (ave) (r,z) r l | r=0 × k=0 l l! k!(lk)! r lk { δ(z) } k cos k 2πft.
l=2m 1 l! l I 1 (ave) (r,z) r l | r=0 l! ( 2m )!(l2m)! r l2m { δ(z) } 2m cos2π2mft 2 2m1 = l=2m 1 (l2m)! l2m r l2m ( 2m I 1 (ave) (r,z) r 2m ) | r=0 r l2m × 1 2 2m1 ( 2m )! δ (z) 2m cos2π2mft = 2m I 1 (ave) (r) r 2m 1 2 2m1 ( 2m )! δ (z) 2m cos2π2mft.
S 2mf (r,z) ( 1 ) m η σ (2) I 2 (ave) (r,z)δ (z) 2m 2m I 1 (ave) (r) r 2m T 2 2m ( 2m )! f L τ .
S 2f (r,z) δ 0 2 (z+ f OB ) 2 2 f OB 2 D 2 (z) ( 1 4 r 2 D (z) 2 ) S conv (r,z),
D(z)= w 0 1+ z 2 / ρ 2
ρ= nπ w 0 2 λ c .

Metrics